Microbiological reduction of 1,3,-dioxo-2-alkylcycloalkanes

ABSTRACT

An improved method of microbiologically reducing a 1,3-dioxo-2alkylcycloalkane using members of a group of compounds which increase the production of the 1 Beta -hydroxy-2 Beta -alkyl-3oxocycloalkane product. The products are useful as intermediates in the total synthesis of useful steroid compounds. Alkyl alcohol and acrylonitrile are illustrated as representative of the useful group of compounds.

United States Patent Lanzilotta Feb. 19, 1974 [54] MICROBIOLOGICALREDUCTION OF 3,579,544 5/1971 Hiraga et a]. 195/51 R 13, DI0XO 2ALKYLCYCLOALKANES 3,616,226 10/1971 Isono et a]. 195/51 R R 211 S v [75]Inventor zg P an Jose Primary Examiner--Alvin E. Tanenholtz Attorney,Agent, or Firm -Evelyn K. Merker [73] Assignee: Syntex Corporation [22]Filed: July 24, 1972 57 ABSTRACT PP 274,297 An improved method ofmicrobiologically reducing a l,3-dioxo-2-alkylcycloalkane using membersof a 52 us. Cl 195/51 R greuP ef eehlP"lmds Whieh h'ereeee thePreduefieh Of [51] Int. Cl ClZb 1/00 the B' y y- B- kyl3-OX0Cycl0alkaneproduct. 58 Field of Search 195/51 R The Predhete are useful asintermediates in the synthesis of useful steroid compounds. Alkylalcohol [56] References Cited and acrylonitrile are illustrated asrepresentative of UNITED STATES PATENTS the useful group of compounds.

3/1969 Bellet et al..-. 195/51 R 16 Claims, No Drawings The presentinvention relates to an improved method of reducing oxo-containingcompounds by microbiological means. In particular, the present inventionis directed to an improved method of effecting the selectivemicrobiological reduction of symmetrical dioxocycloalkanes to thecorresponding optically active oxohydroxycycloalkanes.

The use of microorganisms to effect the reduction of an oxo group hasbeen described, for example, in US. Pat. Nos. 3,432,393, 3,481,974,3,562,112, and 3,616,226.

The reduction of symmetrical 2-alkylcycloalkane- 1,3-diones as describedtherein, e.g., a l3-carbyl-8,l4- secogona-14,l7-dione, results in amixture of products wherein each of the 2-alkyl group and resultant 1-hydroxy group (i.e., the 13-carbyl and resultant l7- hydroxy groups ofthe 8,14-secogona compound) is in the alpha and beta positions. Inasmuchas the reduced products are useful as intermediates for the preparationof pharmacologically active steroid compounds, the optically activelfl,2B(l7B,l3B) reduction product is most useful and therefore desired.However, the prior art methods have not been altogether successful inselectively producing this desirable isomer in reproducibly attractiveyields.

Now it has been discovered that symmetrical dioxocycloalkanes can bemicrobiologically reduced in good and improved yields of the selected113,28 isomer product by the use of certain microbiological speciestogether with members of a group of compounds which unexpectedlyincrease the selective reduction. In accordance herewith, selectivereduction with enhanced production of the lB-hydroxy-ZB-alkyl product iseffected by utilizing microbiological species of the genus Saccharomycesand Schizosaccharomyces genera together with a member of a group ofenhancer compounds. The microbiological species interacts with theenhancer compound to afford increased yields and throughout of thedesired 15,213 reduction isomer.

The present invention thus involves an improvement of the process ofmicrobiologically reducing a 1,3-dioxo-2-alkylcycloalkane whichcomprises employing a microorganism selected from the Saccharomyces andSchizosaccharomyces genera together with a compound selected from thegroup consisting of allyl alcohol, methallyl alcohol, acrylonitrile,methacrylonitrile, acrylic aldehyde, a-methylacrylic aldehyde,a,B-unsaturated ketones, and a-methyl-a,B-unsaturated ketones, saidcompound being employed in an amount sufficient to enhance theproduction of the 1B-hydroxy-2B-alkyl- 3-oxocycloalkane product, andoptionally recovering said product.

Cultures of species of the Genera Saccharomyces and Schizosaccharomyceswhich can be employed in the process of the present invention areavailable from known sources, such as the American Type CultureCollection (ATCC), Rockville, Maryland; the Northern UtilizationResearch and Development Branch, US. Department of Agriculture (NRRL),Peoria, Illinois; and Centralalbureau voor Schirnmelcultures (CBS),Baarn, Holland. The following species are typical of those availablefrom the above sources and are representative of those which can beemployed in the process of the present invention.

Saccharomyces cerevlsleae ATCC 4097 Y-l47 NRRL Saccharomyces uvarum ATCC10609 Y-672 NRRL Saccharomyces pastorianus ATCC 12752 Y-l55l NRRLSchizosaccharomycerjapanlcus ATCC 10660 Y-136l NRRL SchizosaccharomycesCBS 5557 malidevorans Schizosaccharomyces octosporus ATCC 2479 Y-854NRRL ATCC 4206 Schizosaccharomyces pombe ATCC 2476 Y-l64 NRRL ATCC 2478Y-9 NRRL ATCC 14548 ATCC 16491 ATCC 16979 Schizosaccharomyces versatilisATCC 9987 Y-l026 NRRL Of particular importance are the strainsSchizosaccharomyces pombe ATCC 2476 and malidevorans CBS 5557 andSaccharomyces cerevisieae ATCC 4097 and uvarum ATCC 10609.

In addition to allyl alcohol, methallyl alcohol, acrylonitrile,methacrylonitrile, acrylic aldehyde, and a-methylacrylic aldehyde,useful enhancer compounds include a,B-unsaturated ketones anda-methyl-afiunsaturated ketones containing from three to about 12 carbonatoms, including 3-oxo-6-carbomethoxy-n-hexl-ene, cyclohex-1-en-3-one,methyl vinyl ketone, methyl isopropenyl ketone, ethyl vinyl ketone,ethyl isopropenyl ketone, propyl vinyl ketone, propyl isopropenylketone, non-1-en-3-one, dodec-l-en-3-one, and the like. In general,certain of the useful enhancer compounds hereof can be represented bythe following general formula:

' wherein X is hydrogen or methyl, and Y is hydroxywherein,

R is methylene or ethylene;

R is methyl, ethyl, or propyl;

R is a linear group containing three to 18 carbon atoms, inclusive, or acyclic group of the partial formulas: 1

El] VK a -R -s R o. l

wherein each of R, R and R are as defined hereinabovef The process ofthe present invention is particularly useful for preparing, in a firstgroup, lB-hydroxy-ZB- methyl-or -ethyl-3-oxocyclopentanes (D; R ismethylene,'R is methyl or ethyl) and, in a second group, those compoundsof the first group wherein R is a member selected from the groupconsisting of: 2-carboalkyloxyethyl, 3-oxo-n-butyl, 3-oxo-n-pentyl,3-oxo- 6-carboalkyloxy-n-hexy], 3,7-dioxo-n-octanyl,3,7-dioxo-n-nonanyl, and a group of the formula:

wily,

in which R is lower alkyl, cyclopentyl, cyclohexyl, or lower hydrocarboncarboxylic acyl and each of Z, Z

' and Z is a carbon-carbon single bond or a carboncarbon double bond,provided that (1') when Z is a double bond then each of Z and Z is asingle bond and (2) when Z' is a single bond then each of Z and Z is asingle bond or each of Z and Z is a double bond or Z is a double bondand Z is a single bond. Particularly preferred as substrates are2-carboalkyloxyethyl, 3- oxo6-carboalkyloxy-n-hexyl, 2-( 6-methoxy- 1,2,3,4- tetrahydronaphth-2-yl)-ethyl,2-(6-methoxy-3,4-dihydronaphth-2-yl)-ethyl, and 2,2-(6-methoxy-l,2,3,4-tetrahydronaphth-2-ylidene) ethyl.

The lB-hydroxy-2B-alkyl-3-oxocycloalkane products of the process of thepresent invention are useful as intermediates in the preparation ofknown and useful steroid compounds by total synthetic methods known inthe art. By virtue of this utility, the cycloalkane ring is destined asRing D of the steroid nucleus and, therefore, the substituentsrepresented by R and, particularly R are chosen in identity according tothe preparation of known and basic steroid compounds having usefulproperties. The above-cited U.S. patents, as well as numerous otherpatents and literature references, document the known syntheticsequences in which the products hereof are useful. An important aspectin these synthetic methods involves the use of an optically resolvedlB-hydroxy-2B-alkyl-3-oxocycloalkane, as noted supra. The instantinvention .permits the unexpected high yield obtention of theparticularly suitable optical isomer useful for further elaboration inthe synthetic sequence of preparing a steroid nucleus having properstereochemistry, i.e., the 1B,2B-isomer.

In accordance herewith, the microbiological reduction is conducted bycontacting a 1,3-dioxo-2- alkylcycloalkane with a species of the generaSaccharomyces or Schizosaccharomyces together with a member of a classof enhancer compounds. This reduction is conducted at temperaturesranging from about 20 C to about 35 C for a period of time sufficient toproduce reduction, preferably ranging from about 24 hours to abouthours. The reduction is conveniently conducted in a nutrient mediumwhich contains sources of carbon, minerals, nitrogen, and vitamins. Themedia preferably has an adjusted pH level. The incubation is furtherconducted under aerobic conditions.

In carrying out the reduction process, the microorganism enhancercompound, and substrate are contacted andmaintained together in anyorder or fashion convenient and acceptable with conventional microbialfermentation techniques. The reactants are maintained together in asuitable nutrient medium within the given temperature range and for aperiod of time sufficient to produce reduction. Upon the completion ofthe reduction, the mixture is processed by conventional manners toisolate and recover the product. Such manners include filtration,decantation, extraction, evaporation, chromatography, and so forth.

As sources for carbon are employed, the various known and usuallyemployed materials high in carbohydrates such as sugars, starches, andlower organic hydrocarbons.

Minerals are supplied by the addition of inorganic salts. Sufficientamounts of minerals are often present in the other ingredients making upthe fermentation medium for the present process.

As nitrogen sources there can be employed, for example, peptone, soyabean meal, corn steep liquor, yeast extract, caseine, hydrolyzate, andthe like.

Vitamin sources generally include those sources useful in supplying theother necessary nutrients. Yeast extract is especially useful as avitamin source.

The pH of the employed medium is preferably ad-, justed to about 3.5 toabout 4.5 conveniently with mineral acids, e.g., hydrochloric acid.

For the purposes of the present invention, the term an amount sufficientto increase the production of the lB-hydroxy-2B-alkyl-3-oxocycloalkaneproduct will vary depending upon particular substrate choice andenhancer compound choice. In general, enhancer compound is employed inan amount sufficient to provide a ratio of enhancer compound tosubstrate of from about H000 to about 1:10, by weight.

The process of the present invention can be carried out conveniently byfirst cultivating the microorganism in a suitable medium containingadequate carbon, mineral, nitrogen, and vitamin nutrients. Thecultivating medium is conveniently kept under sterile, aerobicconditions and at from about 20 C to about 35 C for from about 12 toabout 48 hours. During this time, the

medium is conveniently agitated.

Following the cultivation period, the culture is transferred, preferablyunder sterile conditions, to a medium of the same or different nutrientcomposition for incubation. The culture is preferably allowed to furthergrow in this medium for from about 8 hours to about 20 hours. After thisadditional growth period, if employed, the substrate and the enhancercompound are added and the resultant mixture is agitated under aerobicconditions within the given temperature and time ranges. The enhancercompound can be added in a single total amount or it can be added to themedium by means of multiple additions of fractions of the total amountemployed and over the whole or part of the incubation reduction. Afterthis time, the product can be isolated by column chromatography with anadequate eluant or by other conventional means.

In one embodiment, the culture is transferred from agar slants, uponwhich it is maintained, to a prepared nutrient medium as defined above.The culture is allowed to grow in this medium at about the sametemperature and preferably with agitation. After the growth period, theculture is used as inoculum for a nutrient medium of the same ordifferent composition. The culture is preferably allowed to grow furtherin the new medium after inoculation.

After the culture has attained a satisfactory stage of growth, thesubstrate, conveniently together with enhancer compound, is added to themedium, preferably as a solution in organic solvent, and the incubationis allowed to proceed. Suitable solvents include acetone, methanol,ethanol, propanol, dioxane, tetrahydrofuran, dimethylsulfoxide, dimethylformamide, and the like, or mixtures thereof. Substrate dissimilationabd product formation can be followed through the use of thinlayer orpaper chromatography of aliquot samples. Column chromatography ispreferably employed for the separation and purification of product atreaction end.

The following examples further illustrate the manner by which thepresent invention can be practiced and represent, in one aspect, thebest mode for carrying out the invention. As such, however, they shouldnot be construed merely as illustrative and not as limitative upon theoverall scope hereof.

EXAMPLE 1 Cells from a 13 to 14 day culture of Schizosaccharomyces pombe(ATCC 2476) are harvested from malt extract-peptone (MP-3) agar mediumand used to inoculate 250 ml. incubation flasks containing 50 ml.portions of nutrient brothof the following composition:

beef extract-3.0 g.

peptone5.0 g.

cerelose20.0 g.

H O (dist.)--l,000 ml.

The cultures are incubated on a rotary shaker at 23 C for 30 hours.After this period, 5 ml. portions of the culture medium are used toinoculate 50 ml. portions of a separate nutrient broth-cerelose mediumof the same composition. After inoculation, the culture is allowed togrow aerobically for an additional 18 hours. At the endof this period,25 mg. of 1,3-dioxo-2-(3-oxo-6-carbomethoxy-n-hexyl)-2-methylcyclopentane substrate in 0.2 ml. ofacetone and a second 25 mg. amount of substrate in acetone after 4 to 5hours and a third 25 mg. amount of substrate in acetone after to 12hours are added to each medium portion while the incubation is continuedat from 22 C to about 24 C with continued shaking as before.

3-Oxo-6-carbomethoxy-n-hex-l-ene in concentration levels of 0.5 mg. and0.25 mg. per 50 ml. medium portions are also added as acetone solutionsl0 #1. and 20 1., respectively) at the same time as the first additionof the substrate. Two 50 ml. medium portions receive 10 pl. and 20 11.1.of acetone, respectively, but no 3-oxo-6-carbomethoxy-n-hex-l-ene, andserve as controls.

The incubation is allowed to proceed in this manner. Two milliliteraliquots are periodically withdrawn from the inoculum mixture after thelast addition of substrate to determine the extent and rate ofreduction. In this determination, each aliquot is extracted with onehalfvolume of chloroform. The chloroform extracts are separated by thinlayer chromatography on silica gel using an :20 (v/v) solvent system ofchloroform- :acetone and the usual detection method employing a spraywith 3 percent ceric sulfate in 3N sulfuric acid solution and heating toC.

After 24 hours, a chromatogram conducted, as above described, indicatesa greater conversion of substrate to the1B-hydroxy-2a-(3-oxo-6-carbomethoxy-nhexyl)-2,'3-methyl-3-oxocyclopentaneproduct with the media containing 3-oxo-6-carbomethoxy-n-hex-l-ene ascompared with the controls.

After hours, a similar chromatogram shows a more dramatic enhancement ofthe lB-hydroxy-2B- methyl product with very little or no by-product andremaining substrate for the 3-oxo-6-carbomethoxy-nhex-1 -ene treatedsamples while the untreated samples show a relatively poor conversion ofsubstrate to desired lB-hydroxy-2B-methyl product and a greater ratio ofby-product to desired product.

EXAMPLE 2 The procedure of Example 1. is repeated except the compoundslisted below in the designated amounts are added to individual 50 ml.medium portions in lieu of the amounts of3-oxo-6-carbomethoxy-n-hex-l-ene added as recited therein.

Medium Portion No. Compound Added Amount, Mg. (in

Chromatograms taken as described in Example 1 at A 24, 48, and 120 hoursafter the addition of substrate,

show an enhancement of the production of the 1B-hydroxy-2B-methyl-2a-(3-oxo-6-carbornethoxy-nhexyl)-3-oxocyclopenta.neproduct for each of medium EXAMPLE 3 The procedure of Example 1 isrepeated except a total of seven 25 mg. additions of substrate inacetone are made to the 50 ml. medium portions in lieu of the three suchadditions. The first addition is made 18 hours after incubation and theother additions as follows: 5, 12, 24, 32, 48, and 56 hours after thefirst addition.

In the present procedure, one medium portion (No. 1) receives substrateonly and No. 2 receives 0.25 mg. of 3-oxo-6-carbomethoxy-n-hex-l-enewith each addition of substrate.

Chromatograms made 120 hours after the first addition of substrateindicate a greater amount of desired 1,8-hydroxy-2B-methyl product inNo. 2 compared with control.

EXAMPLE 4 The procedure of Example 3 is repeated except allyl alcohol isused in lieu of 3-oxo-6-carbomethoxy-n-hexl-ene and two portions areemployed, one receiving substrate only (control) and the other 0.50 mg.of allyl alcohol with each of the seven additions of 40 mg. each ofsubstrate. ln each portion, each addition after the third is accompaniedby 0.6 ml. of nutrient brothcerelose solution of the composition:

beef extract30.0 g.

peptone-50.0 g.

cerelose200.0 g.

H O (dist.)-1,000 ml.

Chromatograms made 24, 48, and 120 hours after the first addition .ofsubstrate indicate an enhanced conversion to the desiredIB-hydroxy-ZB-methyl-Za- (3 -oxo-6-carbomethoxy-n-hexyl)-3-oxocyclopentane as compared with the control.

EXAMPLE The procedure of Example 4 is repeated except seven pairs ofincubation flasks are used, all containing 50 ml. of nutrient broth and2 percent cerelose medium. Supplement additions of nutrient broth andcerelose are made as described in Example 4. One of the seven pairsreceives allyl alcohol (10 pg/ml. medium) with only the first of sevenadditions of 40 mg. each of substrate. The second pair receives allylalcohol (10 pg/ml. medium/addition) with the first two of sevenadditions of 40 mg. eachof substrate, and so forth up to seven additionsof allyl alcohol with seven additions of substrate at the sameconcentration levels.

Chromatograms are made, as described above, 0, 18, 23, 40, and 64 hoursafter the first addition of substrate. These indicate that dissimilationof substrate and production of the desired IB-hydroxy-ZB-methyl productis greatest with the pair of flasks receiving seven additions of allylalcohol with the respective seven additions of substrate. Also shown isthe increase of substrate dissimilation and production of desired 13-hydroxy-2B-methyl product over the control with increasing numbers ofallyl alcohol additions.

EXAMPLE 6 The procedure of Example 1 is repeated except that 0.50 mg. ofallyl alcohol as enhancer is added with each of the three additions ofsubstrate and the following microorganisms are used in lieu ofSchizosaccharomyces pombe ATCC 2476:

Saccharomyces cerevisieae-ATCC 4097 Saccharomyces uvarum-ATCC 10609Saccharomyces pastorianius-ATCC 12752 Schizosaccharomycesmalidevoraus-CBS 5557 Chromatograms taken at 24 and 48 hours after thefirst addition indicate the enhanced conversion to the desired1,B-hydroxy-2B-methyl-2a-( 3-oxo-6-carbomethoxy-n-hexyl)3-oxocyclopentane product as compared with thecontrol.

EXAMPLE 7 The procedure of Example 5 is repeated using 1,3-dioxo-2-(Z-carbomethoxyethyl)-2-methylcyclopentane to obtain enhancedamounts of the desired 1 B-hydroxy-2a-(2-carbomethoxyethyl)-2B-methyl-3- oxocyclopentane product over thecontrol.

In accordance with the foregoing methods, the following are performed.

By incubating 1,3-dioxo-2-( 3-oxo-n-butyl)-2- ethylcyclohexane withSchizosaccharomyces pombe (ATCC 16491) and allyl alcohol (0.5 mg.) in anutrient medium of the composition of Example 1, there is obtainedenhanced amounts of the 1B-hydroxy-2a-(3-oxo-n-butyl)-2,8-ethy1-3-oxocyclopentane product, compared with controlreceiving no allyl alcohol.

By incubating 1,3-dioxo-2-(3-chlorobut-2-enyl)-2-npropylcyclopentanewith Schizosaccharomyces japonicus (ATCC 10660)'and acrolein (1.25 mg.)in a nutrient medium of the composition of Example 1, there is obtainedenhanced amounts of the 1B-hydroxy-2a-(3-chlorobut-2-enyl)-2B-n-propyl-3-oxocyclopentane product, compared withcontrol receiving no acrolein.

By incubating 3-ethoxy-8,l4-secoestra-l,3,5(10)- triene-l4,l7-dione withSchizosaccharomyces pombe (ATCC 2478) and methyl ethynyl ketone in anutrient medium of the composition of the Example 1, there is obtainedenhanced amounts of the 3-ethoxy-8,14.- secoestra- 1 ,3 ,5-( l0)-trien-l7,8-01-14-one product, compared with control receiving no methyl ethynylketone.

By incubating 1 ,3-dioxo-2-(Zcarbopropoxyethyl)-2- ethylcyclohexane withSchizosaccharomyces pombe (ATCC 14548) and allyl alcohol in a nutrientmedium 1 of the composition of Example 1, there is obtained enhancedamounts of the 1B-hydroxy-2a-(2- carbopropoxyethyl)-2,B-ethy1cyclohexaneproduct, compared with control receiving no allyl alcohol.

Among the products prepared in accordance with the foregoing methods arethe following:

1 B-hydroxy-2a-( 3-oxo-n-butyl )-2B-methyI-3- oxocyclopentane,

l B-hydroxy-2a-( 3-oxo-n-butyl )-2B-ethyl-3- oxocyclopentane,

l B-hydroxy-2a-( 3-oxo-n-butyl )-2B-ethyl-3- oxocyclohexane,

lB-hydroxy-2a-(3-oxo-n-pentyl)-2B-methyl-3- oxocyclopentane,

lB-hydroxy-2a-(3-oxo-6'-carboethoxy-n-hexyl)-2B-methyl-3-oxocyclopentane,

lB-hydroxy-2a-(3-oxo-6'-carboethoxy-n-hexyl)-2B-ethyl-3-oxocyclopentane,

lB-hydroxy-2a-( 3-oxo-6 '-carboethoxy-n-hexyl )-2B-methyl-3-oxocyclohexane,

lB-hydroxy-2a-( carbomethoxyethyl)-2B-methyl-3- oxocyclopentane,

lB-hydroxy-2a-(carboethoxyethyl)-2B-ethyl-3- oxocyclopentane,

lB-hydroxy-2a-( carboethoxyethyl )-2B-ethyl-3- oxocyclohexane,

lB-hydroxy-2a-( 3 ,7-dioxo-n-octanyl )-2B-methyl-3- oxocyclopentane,

lB-hydroxy-2a-( 3 ,7-dioxo-n-octanyl )-2B-methyl-3- oxocyclohexane,

lB-hydroxy-2a-( 3 ,7-dioxo-n-nonanyl )-2B-methyl-3- oxocyclopentane,

lB-hydroxy-2a-( 3 ,7-dioxo-n-nonanyl )-2B-methyl-3- oxocyclohexane, and

lB-hydroxy-2a-( 3 ,7-dioxo-n-octanyl )-2B-ethyl-3- oxocyclopentane.

What is claimed is:

1. Process for microbiologically reducing a l,3-dioxo-2-alkylcycloalkanesubstrate which comprises subjecting said substrate to the action of amicroorganism selected from the Saccharomyces and Schizosaccharomycesgenera in the presence of a compound selected from the group consistingof allyl alcohol, methallyl alcohol, acrylonitrile, methacrylonitrile,acrylic aldehyde, a-methylacrylic aldehyde, and a, B-unsaturated ketonesand a-methyl-a, B-unsaturated ketones containing from three to about 12carbon atoms, said compound being present in an amount sufficient toenhance the production of the lB-hydroxy-2B-alkyl-3- oxocycloalkaneproduct and recovering said product.

2. The process according to claim 1 wherein thel,3-dioxo-2-alkylcycloalkane substrate is one of the formula:

and the corresponding product lB-hydroxy-2B-alkyl-3- oxocycloalkane isof the formula:

wherein,

R is methylene or ethylene, R is methyl, ethyl, or propyl; and R is amember selected from the group consisting of:

2-carboalkyloxyethyl, 3-oxo-n-butyl, 3-oxo-npentyl,3-oxo-6-carboalkyloxy-n-hexyl, 3,7-dioxon-octanyl, 3,7-dioxo-n-nonanyl,and

in which R is a lower alkyl, cyclopentyl, cyclohexyl, or lowerhydrocarbon carboxylic acyl and each of Z Z and Z is a carbon-carbonsingle bond or a carboncarbon double bond, provided that (1) when Z is adouble bond then each of Z and Z is a single bond and (2) when Z is asingle bond then each of Z and Z is a single bond or each of Z and Z isa double bond or Z is a double bond and Z is a single bond.

3. The process according to claim 2 wherein said compound is allylalcohol, acrylonitrile, acrylic aldehyde, methyl vinyl ketone, or3-oxo-6-carbomethoxyn-hexl -ene.

4. The process according to claim 3 wherein the microorganism is of thegenus Saccharomyces.

5. The process according to claim 3 wherein the microorganism is of thegenus Schizosaccharomyees.

6. The process according to claim 2 wherein said compound is allylalcohol, acrylonitrile, or acrylic aldehyde.

7. The process according to claim 6 wherein the microorganism is of thegenus Saccharomyces.

8. The process according to claim 6 wherein the microorganism is of thegenus Schizosaccharomyees.

9. The process according to claim 8 wherein the microorganism isselected from the group consisting of Schizosaccharomyces pombe ATCC2476, Schizosaccharomyces malidevorans CBS 55 57, Saccharomycescerevisieae ATCC 4097, and

Saccharomyces uvarum ATCC 10609.

10. The process according to claim 8 wherein the microorganism isSchizosaccharomyces pombe ATCC 2476.

11. The process according to claim 10 wherein the compound is allylalcohol.

12. The process according to claim 3 wherein R is ethyl.

13. The process according to claim 3 wherein R is methyl.

14. The process according to claim 13 wherein R is3-oxo-6-carbomethoxy-n-hexyl.

15. The process according to claim 14 wherein the microorganism is ofthe genus Schizosaccharomyees.

16. The process according to claim 14 wherein the microorganism isSchizosaccharomyces ATCC 2476 and said compound is allyl alcohol.

2. The process according to claim 1 wherein the1,3-dioxo-2-alkylcycloalkane substrate is one of the formula:
 3. Theprocess according to claim 2 wherein said compound is allyl alcohol,acrylonitrile, acrylic aldehyde, methyl vinyl ketone, or3-oxo-6-carbomethoxy-n-hex-1-ene.
 4. The process according to claim 3wherein the microorganism is of the genus Saccharomyces.
 5. The processaccording to claim 3 wherein the microorganism is of the genusSchizosaccharomyces.
 6. The process according to claim 2 wherein saidcompound is allyl alcohol, acrylonitrile, or acrylic aldehyde.
 7. Theprocess according to claim 6 wherein the microorganism is of the genusSaccharomyces.
 8. The process according to claim 6 wherein themicroorganism is of the genus Schizosaccharomyces.
 9. The processaccording to claim 8 wherein the microorganism is selected from thegroup consisting of Schizosaccharomyces pombe ATCC 2476,Schizosaccharomyces malidevorans CBS 5557, Saccharomyces cerevisieaeATCC 4097, and Saccharomyces uvarum ATCC
 10609. 10. The processaccording to claim 8 wherein the microorganism is Schizosaccharomycespombe ATCC
 2476. 11. The process according to claim 10 wherein thecompound is allyl alcohol.
 12. The process according to claim 3 whereinR1 is ethyl.
 13. The process according to claim 3 wherein R1 is methyl.14. The process according to claim 13 wherein R2 is3-oxo-6-carbomethoxy-n-hexyl.
 15. The process according to claim 14wherein the microorganism is of the genus Schizosaccharomyces.
 16. Theprocess according to claim 14 wherein the microorganism isSchizosaccharomyces ATCC 2476 and said compound is allyl alcohol.